| Co-Investigator(Kenkyū-buntansha) |
TAKAOKA Kunio OSAKA CITY UNIVERSITY, Graduated school of medicine, Professor, 大学院医学研究科, 教授 (30112048)
KAZUKI Kenichi OSAKA CITY UNIVERSITY, Graduated school of medicine, Assistant professor, 大学院医学研究科, 助教授 (80254407)
IWAKI Hiroyoshi OSAKA CITY UNIVERSITY, Graduated school of medicine, Instructor, 大学院医学研究科, 講師 (20381981)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2006: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2005: ¥3,100,000 (Direct Cost: ¥3,100,000)
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| Research Abstract |
First of all, we examined MC3T3-E1 and a ST-2 cell whether they make a change on a signal transmission course by making them stretch in cell extension device ST-140 after exposing them to Rho kinase inhibitor Y27632. But there was no difference in ALP activity by the low density dosage and we could not make them stretch by the high density dosage because cells were detached from chamber. Secondly, we cultured El and ST-2 in a silicon chamber to confirm whether activity of Rho contributed by stretch, and made stretch for 12 hours and compared RhoA activity with control, but we could not recognize the significant difference.
Next, we examined correlation of stretch and BMP signal. At first we examined ALP activity for BMP4 with ST-2 and El, and ST-2 reacted for capacity dependence for BMP, whereas the response of El was bad. Successively, it was recognized that stretch of 1% was a suitable to raise ALP activity for ST-2, however the ALP activity fell adversely if we cultured ST-2 with 5%BMP4 50ng/ml for 48 hours after stretching 1% for 24 hours, so that the synergistic effect was not observed.
As future work, we develop mutant low molecular G protein (dominant negative) and examine correlation with stretching, because a method to use Rho inhibitor for directly obstructs the cell adhesion and inhibits stretching,
In addition, we showed a data the cAMP reinforce action of BMP complementarily in our laboratory. Thus, we measure the cAMP density of a cell by ELISA because the signal suppression of RhoA is expected to be different from MAPK course. On that occasion, we will evaluate correlations with other signal cascades by using inhibitor of them.
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